GSM7926419: Phf6 cKO Committed cells RNA-Seq Rep3; Mus musculus; RNA-Seq
150,000 GFP+ committed leukemic cells were sort purified into TRIzol LS reagent (Thermo Fisher Scientific, 10296010). Total RNA was isolated using RNeasy mini kit (Qiagen 74106) by following manufacturer recommended protocol. RNA concentration was determined by using Agilent 2200 TapeStation and a high sensitivity RNA Screentape (Agilent 5067-5579). Samples with a RIN greater than or equal to 8 were used to construct RNA-Seq libraries. RNA libraries were prepared using Smarter Stranded Total RNA-Seq Kit - Pico Input Mammalian (Takara Bio 635005) using the manufacturer recomended protocol. Concentration and the size of the libraries were detremined using Agilent 2200 TapeStation and KAPA library quantification kit (Roche KK4824). RNA libraries were prepared using NEBNext® Ultra™ II directional RNA library prep kit (New England Biolabs, E7530L) using the manufacturer recommended protocol. Concentrations and sizes of libraries were determined using Agilent 2200 Tapestation and KAPA library quantification kit (Roche KK4824). Libraries from all samples were pooled in equimolar concentrations and denatured along with 1% PhiX for paired-end sequencing. Pooled libraries were quantified using KAPA library quantification kit (Roche KK4824).